This application addresses broad Challenge Area (06) Enabling Technologies and specific Challenge Topic, 06-EB-102, Development of Biomedical Technologies and Systems. Multiplexed diagnostic assays that are rapid, easy to use, and very low cost are needed for use at the point of care (POC), especially in low-resource settings. Microfluidic systems can bring laboratory-quality multiplexed assays to POC applications, but so far they require benchtop "readers" that cost too much for use in most low-resource settings. Lateral flow assays are low cost and easy to use by untrained people, but generally measure only one analyte per device, and are not capable of multi-step chemical processing for sophisticated amplification and sample pretreatment steps, resulting in lower test sensitivity and reproducibility. We propose to combine the sophistication of the microfluidic circuit with the simplicity of the conventional lateral flow assay. We plan to show that a novel diagnostic platform, two-dimensional paper networks (2DPNs), can be used without an instrument to perform complex sets of chemical processes and at a cost-per-test that will be comparable to conventional lateral flow tests. Specifically, we propose to demonstrate a sensitive multiplexed immunoassay using disposable 2DPNs for use in low-resources settings. The platform will have the following capabilities;(1) detection of multiple targets from 2 drops of blood, (2) a time to result of less than 10 minutes, (3) on-board sample preparation, (4) reagents stored dry, (5) usable by an untrained person, (6) improved sensitivity compared to conventional lateral flow assays, and (7) a device size comparable to a credit card with weight less than 100 gm.